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Figure 4 | Journal of Translational Medicine

Figure 4

From: Screening NK-, B- and T-cell phenotype and function in patients suffering from Chronic Fatigue Syndrome

Figure 4

Analysis of CD4 and CD8 T cell activation, proliferative capacity and death. Panel A. Gating strategy to analyze FOXP3 and Ki67 expression. CD4 and CD8 T cells were identified in a CD3+ gate. In CD4 T cells, the expression of FOXP3 and CD25 defined the Treg population, while remaining cells were analyzed for Ki67 expression. In CD8 T cells, CD5 and Ki67 were analyzed in the whole population. Panel B. The entire CD4 and CD8 T cell gates illustrated in Figure 3 were analyzed for the frequency of Treg (CD25+FOX–P3+) cells, or for the expression of the indicated markers. In all cases, data from healthy donors (n = 25, HD) and SFC affected individuals (n = 19, SFC) are shown. Panel C. PBMC from healthy donors (n = 5, HD) and CFS affected individuals (n = 8, CFS) were stained with CFSE and cultured in the presence of a combination of PHA and IL-2 (PHA/IL-2). Data shown are proliferation index of gated CD4 or CD8 T cells calculated from best-fit curves using FlowJo software. Panel D. PBMC from healthy donors (n = 30, HD) and CFS affected individuals (n = 19, CFS) were also cultured for 24 h to assess spontaneous CD4 or CD8 T-cell death using DIOC6 and PI staining. Results show total cell death defined by low DIOC6 fluorescence signal. In all panels, median values (thick lines), interquartile ranges (boxes) and 10–90 percentile values (bars). In all cases, p-values for nonparametric Mann–Whitney comparison are shown.

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